High-performance liquid chromatographic determination of 6-aminopenicillanic acid by postcolumn alkaline degradation

✍ Scribed by Jun Haginaka; Junko Wakai

• Publisher: Elsevier Science
• Year: 1986
• Tongue: English
• Weight: 388 KB
• Volume: 158
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(86)90602-0

✦ Synopsis

A high-performance liquid chromatographic method has been developed for the determination of 6-aminopenicillanic acid in amino acid mixtures and human serum. The separation of 6-aminopenicillanic acid was carried out on a C18 column using sodium heptylsulfonate or tetrabutylammonium bromide as an ion-pairing agent and methanol as an organic mobile phase modifier. Detection was based on a postcolumn reaction with sodium hydroxide, mercury(II) chloride, and ethylenediaminetetraacetic acid disodium salt followed by measurement of ultraviolet absorbance (at 300 nm) of the reaction product(s). The method is quantitative for 6-aminopenicillanic acid concentrations down to 0.1 microgram/ml in human serum samples with a 20-microliter injection. At a concentration of 2 micrograms/ml, the within- and between-run precisions (relative standard deviation) were 1.29-3.91% and 2.30%, respectively.